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Biomedical subjects

H Blum

Publications and source records attributed to H Blum.

At least 37 records · Page 2Linked to original sources
PubMed1996

Reversible injury: creatinine kinase recovery restores bioenergetics and function.

In postischemia hearts, cytoplasmic creatinine kinase (CK) inactivation resulting from toxic oxygen metabolite injury may lead to bioenergetic and mechanical dysfunction. This study determines the relationship between CK activity, mechanical function, and bioenergetics during reperfusion (RP) after a reversible ischemic injury. Rat hearts pretreated after 12 hr without (CTRL) or with myristic acid (MA) underwent 10 min global, 37 degrees C ischemia followed by 10 or 40 min RP while developed pressure (DP) was monitored. Catalase and CK were assayed at preischemia. CK was also assayed at end ischemia and 10 and 40 min RP. 31 P nuclear magnetic resonance spectra assessed changes in phosphocreatinine (PCr) and adenosine triphosphate (ATP) concentration. Preischemic DP was 95 +/- 5 mm Hg. CTRL DP returned to 84 +/- 3 by RP10 and 88 +/- 6 by RP40 while MA hearts recovered fully by RP10 (90 +/- 2). Preischemic catalase activity was significantly increased in MA hearts (1217 +/- 36 U/g left ventricular tissue (LV) vs 1007 +/- 40 U/g LV, P < 0.01, MA vs CTRL). CTRL CK activity fell from 1870 +/- 75 to 1103 +/- 11 U/g LV at RP10, but rose to 1272 +/- 13 by RP40 (P < 0.01, RP10 vs RP40). MA hearts lost no CK activity during RP. By RP10, CTRL PCr/ATP ratio was elevated to 2.2 +/- 0.2 (P < 0.001) from a preischemic level of 1.7 +/- 0.4 and normalized by RP40, while MA hearts had a normal PCr/ATP throughout RP. Reversible RP injury transiently depresses mechanical function. Cytoplasmic CK damage during RP impairs PCr utilization, leading to a PCr overshoot. Functional recovery and metabolic recovery follow return of CK activity. Increased endogenous catalase preserves CK during RP, resulting in normal function and bioenergetics.

Adenosine Triphosphate↗
PubMed1996

The Irma dream, self-analysis, and self-supervision.

The Irma dream has special historical significance. Erikson and others have placed it in historical, social, and cultural context. The manifest dream was elaborated in terms of analytic surface with analysis of form and content, patterns and movement in time and space, etc. There are, however, limits to textual reinterpretations. Further psychobiographic consideration of the Irma dream highlights issues of transference, countertransference and their sources in unconscious conflict and trauma. The Irma dream was initially a secret dream which represented the initiation of a self-analytic and supervisory process. Freud's revealing the dream and imagining the commemoration of the discovery of "the secret of the dream" marked the termination of formal self-analysis within analysis interminable.

Countertransference↗
PubMed1995

The phosphocreatine overshoot occurs independent of myocardial work.

Although the exact mechanism(s) responsible for the phosphocreatine/ATP overshoot have not been completely elucidated, our data demonstrate that the overshoot does not stem from reduced myocardial work, and consequently, reduced utilization of phosphocreatine (PCr). Additionally, we highlight a basic difference in the physiologic responses of skeletal and cardial muscle to work demands. By understanding the bioenergetic derangements which accompany reperfusion injury, one may hope to better salvage post-ischemic myocardium.

Adenosine Triphosphate↗
PubMed1994

Intracellular sodium in cardiomyocytes using 23Na nuclear magnetic resonance.

Intracellular sodium content in superfused isolated rat cardiomyocytes was measured using 23Na nuclear magnetic resonance. The shift reagent dysprosium tripolyphosphate was added to the buffer to distinguish between NMR signals from the intracellular region and the extracellular buffer. The NMR visibility of the intracellular sodium signal was experimentally determined by measuring the changes induced in the sodium NMR signals by application of ischemia as an intervention. Intracellular volume was accounted for by determining the change in the sodium signal upon adding cells (in beads) to the buffer solution at the beginning of each experiment and by killing the cells (in beads) with Triton X-100 at the end of each experiment. The visibility of intracellular sodium (relative to extracellular) was 0.47 +/- 0.12 (mean +/- S.D., n = 12). The average intracellular sodium concentration using this visibility is 29 +/- 4.5 mM (n = 12). This value is much higher than results obtained by some investigators using NMR techniques and by others using different standard methods, with the exception of those methods which evaluate the total intracellular sodium (atomic absorption spectroscopy and X-ray microanalysis). We conclude that total Nai is higher than generally reported, using other accepted techniques such as ion-specific electrodes, and that 23Na-NMR analysis can be used to accurately determine Nai in intact cells.

Animals↗
PubMed1994

Ischemic preconditioning preserves end-ischemic ATP, enhancing functional recovery and coronary flow during reperfusion.

The mechanisms by which ischemic preconditioning (IPC) protects against reperfusion (RP) injury are unknown. The purpose of this study was to relate IPC to postischemic mechanical function, vascular reactivity, and bioenergetics. Isolated perfused rat hearts were randomized to two groups. Control (CTRL) hearts underwent 25 min of global, 37 degrees C ischemia and 40 min RP. IPC hearts underwent 2.5 min ischemia and 10 min RP followed by 25 min ischemia and 40 min RP (RP40). Left ventricular developed pressure (DP) and coronary flow were continuously measured. 31P NMR spectra determined phosphocreatine and ATP concentrations in parallel hearts every 5 min. Results are means +/- SEM; n = 6/group. Significance was assumed for P < 0.05 by paired (within groups) and unpaired (between groups) t test. CTRL heart DP recovered to 35 +/- 4% of preischemic (PI) DP by RP40 (P < 0.001), while IPC heart DP reached 99 +/- 4% (P = NS vs PI; P < 0.001 vs CTRL). CTRL coronary flow recovered to only 75 +/- 3% of PI (P < 0.001) by RP40. IPC coronary flow exceeded baseline during RP (RP40 = 118 +/- 3%, P < 0.001 vs CTRL; P < 0.05 vs PI). After 25 min ischemia, CTRL heart ATP fell to 40 +/- 4% of PI (P < 0.001) while the IPC group fell to only 60 +/- 4% (P < 0.05 IPC vs CTRL; P < 0.001 vs PI). IPC preserves more end-ischemic ATP compared to CTRL hearts with a resultant improvement in mechanical function during reperfusion. Only preconditioned hearts preserve the adaptive mechanism(s) responsible for postischemic vasodilatation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1994

Hepatic phosphate trapping, decreased ATP, and increased feeding after 2,5-anhydro-D-mannitol.

The mechanism by which the fructose analogue 2,5-anhydro-D-mannitol (2,5-AM) elicits feeding behavior was investigated by studying its metabolism and biochemical effects in liver. Thin-layer chromatography of liver extracts from rats given 2,5-AM containing 14C-labeled 2,5-AM showed that the analogue is phosphorylated in vivo with a time course that parallels the eating response. In vivo 31P nuclear magnetic resonance spectroscopy of rat liver during intravenous infusion of 2,5-AM and high-resolution nuclear magnetic resonance analyses of liver extracts showed that 2,5-AM is rapidly phosphorylated in liver, trapping hepatic phosphate and decreasing ATP, inorganic phosphate, and phosphate diesters. These changes occurred in a time frame in which the feeding response is elicited in conscious animals given the same dose of 2,5-AM by the same route. During an interval in which 2,5-AM increased eating, it also increased urinary uric acid excretion, implicating enhanced adenosine degradation in the reduction in hepatic ATP. These results provide the first direct evidence that changes in a high-energy phosphate-carrying compound in liver may provide a signal to initiate eating behavior.

Adenosine Triphosphate↗
PubMed1993

Protecting myocardial creatine kinase activity during reperfusion improves bioenergetics and contractile function.

The exact mechanisms underlying reperfusion (RP) injury are unclear but are thought to involve toxic oxygen metabolites such as H2O2. The purpose of our study was to determine whether increasing endogenous catalase (CAT) stores would protect creatine kinase (CK) activity and improve bioenergetics and mechanical function during RP. Rats (n = 6/group) were pretreated with myristic acid (MA) or nothing 6 to 12 hr prior to cardiectomy. Hearts were Langendorff perfused and developed pressure (DP) was monitored during 25 min of 37 degrees C ischemia and 10 or 40 min of RP. CK activity was determined at baseline, end ischemia, and 40 min RP. CAT activity and H2O2 production was assayed at baseline, end ischemia, and 10 min RP. 31P NMR spectra were continuously acquired to determine ATP and phosphocreatine (PCr) concentrations. MA-pretreated hearts demonstrated elevated CAT stores (121 +/- 4%, P < 0.05). No H2O2 was produced during ischemia, and both groups generated significant but equal amounts of H2O2 at RP 10 (P < 0.001 vs preischemia (PI), P = NS between groups). By RP 40, MA-pretreated hearts recovered more DP than did control hearts (75 +/- 5% of PI vs 35 +/- 4% of PI, P < 0.001) and retained more CK activity as well (66 +/- 4% vs 52 +/- 2%, P < .05). PCr/ATP ratios of control hearts were abnormally elevated above baseline and that of MA hearts during RP (2.4 +/- 0.1 vs 1.85 +/- 0.08, P < 0.05 at RP 40).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1993

Differential regulation of IL-9-expression after infection with Leishmania major in susceptible and resistant mice.

IL-9 is a pleiotropic lymphokine, one of its activities being the growth stimulation of certain CD4+ T lymphocytes. In murine cutaneous leishmaniasis, depending on the genetic background of the host mouse strain, vigorous proliferation of either mainly Th1 in resistant C57BL/6 mice or Th2-type CD4+ T cells in susceptible BALB/c mice occurs after infection with Leishmania major (L. major). Since little is known about the involvement of IL-9, the possible role of this cytokine with regard to its immunregulatory function was evaluated by comparing its presence in the serum and its expression kinetics in spleen and lymph nodes in resistant and susceptible mice. To this sera of L. major-infected mice were tested functionally for IL-9. In addition the PCR-aided detection of IL-9 mRNA in organs of mice and measurement of the lymphokine in supernatants of restimulated lymph node and spleen cell cultures were used. We show here that although no functionally active IL-9 was detected in sera of both BALB/c and C57BL/6 mice, IL-9 is produced after in vitro antigenic restimulation and its mRNA was found to be expressed in lymph nodes and spleens during an immune response against L. major. Shortly after infection no principal differences in the kinetics of IL-9 expression could be observed, which had its maximum between day 5 and 7 after infection. The rate of production however was higher in the susceptible BALB/c mice. In athymic BALB/c nu/nu mice and in mice depleted of CD4+ T cells no IL-9 production was detectable in vivo at the level of mRNA and no IL-9 was produced after stimulation with L. major antigen in vitro. Treatment of infected mice with cyclosporin A ablates antigen-specific IL-9 production when tested in vitro without affecting its production after polyclonal T cell stimulation. Positively selected, purified CD4+ T cells were fully capable of producing IL-9. From 4 weeks after infection, IL-9 synthesis was observed only in BALB/c mice, correlating with the expansion of antigen-specific Th2 type T helper cells in these mice. Treatment of BALB/c mice with neutralizing anti-IL-4 mAb, a regimen known to lead to subsequent cure of infected BALB/c mice, suppressed late IL-9 synthesis.

Animals↗
PubMed1993

Effect of temperature and coronary flow on the metabolic and mechanical function of the isolated rat heart.

A number of cardiac metabolic intermediates, namely, adenosine triphosphate (ATP), H+, phosphocreatine (PCr), inorganic phosphate (Pi), adenosine diphosphate (ADP), and related functions of these intermediates, Gibbs' free energy of ATP hydrolysis (delta G) and phosphorylation ratio [ATP/(ADP.Pi)], are thought to adjust mitochondrial oxidative phosphorylation rates to conform to mechanical demand. The effects of hypothermia and altered perfusion pressure on these parameters were evaluated in 12 hearts from Sprague-Dawley rats perfused in the Langendorff mode. 31P-nuclear magnetic resonance (NMR) spectra were obtained at cardiac temperatures between 20 and 37 degrees C, and coronary perfusion pressures between 20 and 145 cm H2O. Coronary flow varied between 0.5 and 15 ml/min throughout this range of intervention. Heart rate (HR), left ventricular systolic pressure (LVSP), and specific volumetric coronary flow (SCF) were determined for each temperature and perfusion pressure. The product HR x LVSP directly correlated with perfusion pressure at all temperatures. The temperature dependence could be represented by an overall activation energy of 72.7 kJ/M. In the constant temperature experiment, SCF and HR x LVSP fell linearly with decreasing perfusion pressure. Quantitative evaluation of the relationship between cardiac function and the metabolic intermediates described above defined these intermediates as nonregulatory with the possible exception of H+.

Acid-Base Equilibrium↗
PubMed1992

Isolated cardiomyocytes in conjunction with NMR spectroscopy techniques to study metabolism and ion flux.

To distinguish cellular from vascular responses to physiological and pathophysiological stimuli, we developed methods to perform NMR spectroscopy on isolated ventricular cardiomyocytes. Isolated adult rat cardiomyocytes, placed in agarose beads and superfused with phosphate-free buffer (Media 199 (GIBCO 400-1100) gassed with 95% O2, 5% CO2), were used to evaluate a variety of cellular processes during different pharmacological and physiological interventions. Bioenergetic function was monitored with 31P NMR. Intermediary metabolism, gluconeogenesis, and glycolysis were monitored with 13C NMR. Sodium flux was monitored with 23Na NMR. Calcium flux was monitored with 19F NMR in conjunction with an intracellular calcium-chelating agent, 5F-1,2-bis(2-amino-phenoxy)ethane-N,N,N',N'-tetraacetic acid. Creatine kinase kinetics (forward rate constant (Kf) and flux of phosphocreatine to ATP) were estimated with 31P NMR saturation transfer data. Various combinations of NMR parameters were monitored simultaneously so that the interaction of metabolism and ion flux could be evaluated. We have demonstrated that it is possible to simultaneously monitor a variety of cellular processes in intact heart cells in real time, without the confounding influences of perfusion, contractile function, and extrinsic blood-borne neurohumoral agents. This model will be useful for longitudinal studies of myocyte metabolism and ion flux.

Acetylcholine↗
PubMed1991

Coupled in vivo activity of creatine phosphokinase and the membrane-bound (Na+,K+)-ATPase in the resting and stimulated electric organ of the electric fish Narcine brasiliensis.

Physiological control of the plasma membrane sodium pump, (Na+,K+)-ATPase, is essential for proper function of eukaryotic cells. In the electric organ of the elasmobranch Narcine brasiliensis, the normal demands placed upon the pump during the process of generation of electrical currents call for large and rapid changes in activity of this enzyme, making this a good model for the study of its cellular regulation. 31P NMR spectroscopic techniques were used to study metabolic regulation of membrane pump function in resting and stimulated electric organ and in skeletal muscle of the live, intact N. brasiliensis. Because the ATP synthetic abilities of the electric organ by glycolysis or oxidative phosphorylation are extremely limited, depletion of phosphocreatinine (PCr) could be used to determine the activity of the (Na+,K+)-ATPase after the electric organ was stimulated to discharge, and to measure the net flux from PCr to ATP through the creatine phosphokinase (CPK) reaction in the electric organ. Saturation transfer, an NMR technique which measures exchange rates, was applied to determine the unidirectional flux in the forward direction through the same reaction in the electric organ and in skeletal muscle as a control. The pseudo first-order rate constant kf for the CPK reaction at 24 degrees C in resting electric organ was 0.000 +/- 0.002 s-1 (n = 10) and in skeletal muscle was 0.08 +/- 0.03 s-1 (n = 3). The results demonstrate that in resting electric organ, which is well supplied with CPK, there was no measurable flux through this reaction, although CPK when extracted is highly active. Measured and calculated levels of all substrates for the creatine kinase reaction in the electric organ are similar to those in unstimulated skeletal muscle, where the creatine phosphokinase reaction rates are high in vivo. In contrast to the resting electric organ, during stimulation of the electric organ the measured net rate constant was greater than 0.08 s-1. In addition, as shown by lack of PCr depletion, there was virtually no net turnover of ATP in the resting organ compared to the stimulated organ. The marked difference in the (Na+,K+)-ATPase activity in the resting and activated electric organ confirmed earlier results (Blum, H., Nioka, S., and Johnson, R. G., Jr. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 1247-1251). Together, these results suggest that there is a novel method of coordinate regulation of cellular enzymes of great sensitivity and rapidity.

Adenosine Triphosphate↗
PubMed1991

Sodium flux and bioenergetics in the ischemic rat liver.

Concurrent 23Na and 31P nuclear magnetic resonance spectroscopy has been employed to study the effects of ischemia upon the high-energy phosphagens and sodium ion concentration within the in vivo rat liver. High-energy phosphates in the form of ATP were depleted within 10 min of the onset of ischemia when measured by NMR. However, similar liver samples subjected to analytical biochemistry retained 27 +/- 12% of their ATP after a similar 10-min ischemic insult. Time-dependent 23Na NMR measurements, obtained in the presence of the shift reagent Dy(TTHA) to distinguish intracellular from extracellular sodium, revealed a rapid rise in the intracellular sodium when the liver was made ischemic. Intracellular and extracellular sodium concentrations approached equilibrium with an exponential time constant of 14.7 +/- 7 min. The initial rate of sodium influx was calculated to be 1.50 meq.l-1.min-1. The results indicate that the ischemic liver has a high passive sodium permeability and that NMR detectable 31P signals reflect the actual availability of cytosolic high-energy phosphates to enzymes, in this instance the membrane-bound [Na+, K+]-ATPase.

Adenosine Triphosphate↗
PubMed1991

In vivo mechanisms of myocardial functional stability during physiological interventions.

Metabolic regulatory mechanisms are designed to maintain stable myocardial function during extremes in physiological insult; they can now be studied in vivo and may provide insight into mechanisms of altered myocardial functional decompensation during disease processes. To determine mechanisms of myocardial stability during hypoxia and acute pressure loading, creatine kinase (CK) kinetics (forward rate constant, Kf, and flux of phosphocreatine, PCr, to adenosine triphosphate, ATP), and nicotinamide adenine dinucleotide (NADH) redox state were determined with 31P nuclear magnetic resonance (NMR) and NADH fluorometry, respectively, and correlated with heart work (heart rate x systolic blood pressure, HR x SBP), cardiac output (CO) and O2 consumption (MVO2) in 15 anesthetized open chest dogs. Hypoxia (PaO2 of 30-35 mm Hg) was produced in 6 dogs with an inspired O2/N2 of 200/3,000. Cardiac loading was produced in 9 dogs by administration of norepinephrine (NE, 1 micrograms/kg/min). Each dog acted as its own control. Baseline NADH fluorometry, 31P-NMR saturation transfer and cardiac function measurements were performed simultaneously in each dog, after which the experimental interventions were made. Similar increases in HR x SBP, CO, and MVO2 which occurred during both interventions were associated with different bioenergetic responses. During NE infusion, the Kf of CK increased from control; during hypoxia, the Kf decreased from control (p less than 0.05). Flux of PCr----ATP was significantly lower during hypoxia than during NE infusion (p less than 0.05). PCr was decreased significantly during NE infusion (p less than 0.05). In addition, NADH redox state increased (from baseline of 100%) during hypoxia (140 +/- 10%) and decreased during NE infusion (78 +/- 6%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1990

Activation of the Na+, K(+)-ATPase in Narcine brasiliensis.

The in vivo activation and turnover rates of the sodium pump (Na+, K(+)-ATPase) were investigated in the electrocytes of the electric organ of the elasmobranch Narcine brasiliensis. The Narcine electric organ appears to be an excellent model for the study of sodium pump activation in an excitable tissue. The sodium transmembrane gradient and high-energy phosphagens were concurrently measured by 23Na and 31P NMR spectroscopy. The resting electric organ, which depends primarily on anaerobic metabolism, displays a high concentration of phosphocreatine (PCr). It has an intracellular sodium concentration ([Na+]i) of 20 +/- 10 milliequivalents/liter as estimated by NMR. Electrical stimulation of the nerves innervating the electric organ results in an increase in [Na+]i in the electrolyte and rapid depletion of PCr. Ouabain causes an 85% decrease in utilization of high-energy phosphagens, indicating that rapid PCr turnover in this tissue is mainly due to Na+, K(+)-ATPase activity. From these data we can determine that the rate of sodium pump turnover increases by greater than 3 orders of magnitude within several hundred milliseconds. In excised unstimulated electric organ slices, changes in [Na+]i equivalent to those occurring with stimulation, but induced by hyperosmolar conditions, do not result in increased PCr hydrolysis. We conclude that cholinergic stimulation of the electric organ causes a rapid and extremely large increase in sodium pump turnover, which is regulated predominantly by factors other than [Na+]i.

Animals↗
PubMed1989

The expression of the TMV-specific 30-kDa protein in tobacco protoplasts is strongly and selectively enhanced by actinomycin.

The TMV-encoded 30-kDa protein has been implicated in the cell-to-cell transport of TMV in the infected plant. The polyethylene glycol-mediated inoculation of tobacco protoplasts with TMV particles and TMV RNA was used to compare the time courses of the viral 30-kDa protein synthesis in vivo. Upon infection of protoplasts with TMV RNA, the synthesis of the viral 30-kDa protein starts after 4 to 6 hr, has its maximum after 8 to 10 hr, and decreases. After inoculation of protoplasts with TMV, however, the start of the viral 30-kDa protein synthesis and its maximum are delayed by 2 hr, followed by the same decrease. We show that actinomycin D dramatically stimulates the synthesis of the 30-kDa protein by up to 2 orders of magnitude, whereas the synthesis of the viral 126 kDa, the 183 kDa, and the coat protein is increased only by a factor of 2. Surprisingly, actinomycin V is twice as active as actinomycin D, whereas actinomycin I is nearly inactive. The specific stimulation of the 30-kDa synthesis by actinomycin D in vivo depends neither on the Nicotiana variety nor on the TMV strain used. Final evidence that the 30-kDa protein is truly TMV-derived is provided by the slightly different electrophoretic mobilities of the 30-kDa proteins encoded by TMV strains vulgare, dahlemense, and U2. The identification of the 30-kDa protein in two-dimensional gels was achieved for the first time by a combination of ionic and nonionic detergents for the solubilization of the 30-kDa protein and by the specific stimulation of its synthesis by actinomycin D. The mechanism of the strong and selective actinomycin effect on the viral 30-kDa protein synthesis in vivo is as yet obscure. Actinomycin does not appear to act directly on viral protein biosynthesis, since it neither stimulates the 30-kDa synthesis upon translation of TMV RNA in vitro nor alters the ratio of the products. Actinomycin may rather act by inhibiting selectively the synthesis of a host factor whose synthesis starts at least 4 hr after TMV infection and which strongly inhibits the expression of the viral 30-kDa transport protein.

Capsid Proteins↗